Hõrak R, Kivisaar M
Department of Genetics, Estonian Biocentre, Tartu University, 51010 Tartu, Estonia.
J Bacteriol. 1999 Oct;181(20):6312-8. doi: 10.1128/JB.181.20.6312-6318.1999.
Transposition is a DNA reorganization reaction potentially deleterious for the host. The frequency of transposition is limited by the amount of transposase. Therefore, strict regulation of a transposase is required to keep control over the destructive multiplication of the mobile element. We have shown previously that the expression of the transposase (tnpA) of the Pseudomonas putida PaW85 transposon Tn4652 is positively affected by integration host factor. Here, we present evidence that the amount of the transposase of Tn4652 in P. putida cells is controlled by the transposon-encoded protein (TnpC). Sequence analysis of the 120-amino-acid-long TnpC, coded just downstream of the tnpA gene, showed that it has remarkable similarity to the putative polypeptide encoded by the mercury resistance transposon Tn5041. As determined by quantitative Western blot analysis, the abundance of TnpA was reduced up to 10-fold in the intact tnpC background. In vivo experiments using transcriptional and translational fusions of the tnpA gene and the reporter gene gusA indicated that TnpC operates in the regulation of the transposase of Tn4652 at the post-transcriptional level.
转座是一种对宿主可能有害的DNA重组反应。转座频率受转座酶数量的限制。因此,需要对转座酶进行严格调控,以控制移动元件的破坏性增殖。我们之前已经表明,恶臭假单胞菌PaW85转座子Tn4652的转座酶(tnpA)的表达受到整合宿主因子的正向影响。在此,我们提供证据表明,恶臭假单胞菌细胞中Tn4652转座酶的量受转座子编码蛋白(TnpC)的控制。在tnpA基因下游编码的120个氨基酸长的TnpC的序列分析表明,它与汞抗性转座子Tn5041编码的假定多肽具有显著相似性。通过定量蛋白质免疫印迹分析确定,在完整的tnpC背景下,TnpA的丰度降低了10倍。使用tnpA基因和报告基因gusA的转录和翻译融合进行的体内实验表明,TnpC在转录后水平对Tn4652转座酶进行调控。